Disposable biosensor based on graphene oxide conjugated with tyrosinase assembled gold nanoparticles.

A highly efficient enzyme-based screen printed electrode (SPE) was obtained by using covalent attachment between 1-pyrenebutanoic acid, succinimidyl ester (PASE) adsorbing on the graphene oxide (GO) sheets and amines of tyrosinase-protected gold nanoparticles (Tyr-Au). Herein, the bi-functional molecule PASE was assembled onto GO sheets. Subsequently, the Tyr-Au was immobilized on the PASE-GO sheets forming a biocompatible nanocomposite, which was further coated onto the working electrode surface of the SPE. The characterization of obtained nanocomposite and modified SPE surface was investigated by atomic force microscopy (AFM), transmission electron microscopy (TEM) and scanning electron microscopy (SEM). Attributing to the synergistic effect of GO-Au integration and the good biocompatibility of the hybrid-material, the fabricated disposable biosensor (Tyr-Au/PASE-GO/SPE) exhibited a rapid amperometric response (less than 6s) with a high sensitivity and good storage stability for monitoring catechol. This method shows a good linearity in the range from 8.3×10(-8) to 2.3×10(-5) M for catechol with a squared correlation coefficient of 0.9980, a quantitation limit of 8.2×10(-8) M (S/N=10) and a detection limit of 2.4×10(-8) M (S/N=3). The Michaelis-Menten constant was measured to be 0.027 mM. This disposable tyrosinase biosensor could offer a great potential for rapid, cost-effective and on-field analysis of phenolic compounds.

[1]  Huafeng Yang,et al.  The synthesis of perylene-coated graphene sheets decorated with Au nanoparticles and its electrocatalysis toward oxygen reduction , 2009 .

[2]  Jonathan S. Dordick,et al.  Protein‐Directed Formation of Silver Nanoparticles on Carbon Nanotubes , 2007 .

[3]  Yang Li,et al.  Simultaneous determination of cadmium(II), lead(II) and copper(II) by using a screen-printed electrode modified with mercury nano-droplets , 2010 .

[4]  Chunzhong Li,et al.  Electrocatalytic Oxidation of Glucose by the Glucose Oxidase Immobilized in Graphene‐Au‐Nafion Biocomposite , 2010 .

[5]  Yiyu Feng,et al.  Preparation of a graphene oxide–phthalocyanine hybrid through strong π–π interactions , 2010 .

[6]  C. N. R. Rao,et al.  Covalent and Noncovalent Functionalization and Solubilization of Graphene , 2009 .

[7]  Charlotte Carlsson,et al.  Amperometric screen-printed biosensor arrays with co-immobilised oxidoreductases and cholinesterases , 2005 .

[8]  J. Emnéus,et al.  Amperometric sensors based on tyrosinase-modified screen-printed arrays. , 2003, Talanta.

[9]  Hongyuan Chen,et al.  Direct Electrochemistry and Bioelectrocatalysis of Microperoxidase-11 Immobilized on Chitosan-Graphene Nanocomposite , 2010 .

[10]  Craig E. Banks,et al.  Screen printed electrochemical platforms for pH sensing. , 2009, Analytical methods : advancing methods and applications.

[11]  Yong Liu,et al.  Biocompatible graphene oxide-based glucose biosensors. , 2010, Langmuir : the ACS journal of surfaces and colloids.

[12]  Klaus-Dieter Vorlop,et al.  Methylphenazonium-modified enzyme sensor based on polymer thick films for subnanomolar detection of phenols , 1995 .

[13]  Guodong Liu,et al.  Amperometric tyrosinase biosensor based on Fe3O4 nanoparticles-chitosan nanocomposite. , 2008, Biosensors & bioelectronics.

[14]  Jörg Reichert,et al.  Controlled assembly of protein-protected gold nanoparticles on noncovalent functionalized carbon nanotubes , 2010 .

[15]  Roberto Pilloton,et al.  A disposable Laccase-Tyrosinase based biosensor for amperometric detection of phenolic compounds in must and wine , 2010 .

[16]  Daohong Wu,et al.  A novel tyrosinase biosensor based on biofunctional ZnO nanorod microarrays on the nanocrystalline diamond electrode for detection of phenolic compounds. , 2009, Bioelectrochemistry.

[17]  Guo-Li Shen,et al.  A Mediator-Free Tyrosinase Biosensor Based on ZnO Sol-Gel Matrix , 2005 .

[18]  Zhaoqiang Wu Synthesis and characterization of active ester‐functionalized fluorescent polymers: New materials for protein conjugation , 2008 .

[19]  Qingji Xie,et al.  Biofuel cell and phenolic biosensor based on acid-resistant laccase-glutaraldehyde functionalized chitosan-multiwalled carbon nanotubes nanocomposite film. , 2009, Biosensors & bioelectronics.

[20]  Chun Li,et al.  Flexible graphene films via the filtration of water-soluble noncovalent functionalized graphene sheets. , 2008, Journal of the American Chemical Society.

[21]  L. Mita,et al.  A thionine-modified carbon paste amperometric biosensor for catechol and bisphenol A determination. , 2010, Biosensors & bioelectronics.

[22]  H. Dai,et al.  Noncovalent sidewall functionalization of single-walled carbon nanotubes for protein immobilization. , 2001, Journal of the American Chemical Society.

[23]  Serge Cosnier,et al.  Calcium carbonate nanoparticles: a host matrix for the construction of highly sensitive amperometric phenol biosensor. , 2007, Biosensors & bioelectronics.

[24]  C. McNeil,et al.  Disposable tyrosinase-peroxidase bi-enzyme sensor for amperometric detection of phenols. , 2002, Biosensors & bioelectronics.

[25]  Sandip Niyogi,et al.  Solution properties of graphite and graphene. , 2006, Journal of the American Chemical Society.

[26]  Jue Lu,et al.  Nanometal-decorated exfoliated graphite nanoplatelet based glucose biosensors with high sensitivity and fast response. , 2008, ACS nano.

[27]  Yuyan Shao,et al.  Graphene Based Electrochemical Sensors and Biosensors: A Review , 2010 .

[28]  Yongyan Tan,et al.  Amperometric catechol biosensor based on polyaniline-polyphenol oxidase. , 2010, Biosensors & bioelectronics.

[29]  Erkang Wang,et al.  Self-assembly of cationic polyelectrolyte-functionalized graphene nanosheets and gold nanoparticles: a two-dimensional heterostructure for hydrogen peroxide sensing. , 2010, Langmuir : the ACS journal of surfaces and colloids.

[30]  Xin Wang,et al.  Effect of graphene oxide on the properties of its composite with polyaniline. , 2010, ACS applied materials & interfaces.

[31]  John Silcox,et al.  Atomic and electronic structure of graphene-oxide. , 2009, Nano letters.

[32]  Zhuang Liu,et al.  Nano-graphene oxide for cellular imaging and drug delivery , 2008, Nano research.

[33]  Dermot Diamond,et al.  Development of a biosensor for endocrine disrupting compounds based on tyrosinase entrapped within a poly(thionine) film. , 2004, Biosensors & bioelectronics.

[34]  Hua Bai,et al.  Preparation of Gold Nanoparticle/Graphene Composites with Controlled Weight Contents and Their Application in Biosensors , 2010 .

[35]  P. J. Ollivier,et al.  Layer-by-Layer Assembly of Ultrathin Composite Films from Micron-Sized Graphite Oxide Sheets and Polycations , 1999 .

[36]  T. Kuwana,et al.  Electrochemical stability of catechols with a pyrene side chain strongly adsorbed on graphite electrodes for catalytic oxidation of dihydronicotinamide adenine dinucleotide , 1983 .

[37]  S. Mannino,et al.  Nanofibrous membrane based tyrosinase-biosensor for the detection of phenolic compounds. , 2010, Analytica chimica acta.

[38]  Huaiguo Xue,et al.  A highly stable biosensor for phenols prepared by immobilizing polyphenol oxidase into polyaniline-polyacrylonitrile composite matrix. , 2002, Talanta.

[39]  J. Kochana,et al.  Titania sol–gel-derived tyrosinase-based amperometric biosensor for determination of phenolic compounds in water samples. Examination of interference effects , 2008, Analytical and bioanalytical chemistry.

[40]  H. Jiang,et al.  Controlled hybrid nanostructures through protein-mediated noncovalent functionalization of carbon nanotubes. , 2007, Angewandte Chemie.

[41]  Huafeng Yang,et al.  Graphene/AuNPs/chitosan nanocomposites film for glucose biosensing. , 2010, Biosensors & bioelectronics.

[42]  Hui Liu,et al.  Graphene oxide as a matrix for enzyme immobilization. , 2010, Langmuir : the ACS journal of surfaces and colloids.

[43]  P. He,et al.  Electrochemistry and Electrocatalysis of Hemoglobin on 1-Pyrenebutanoic Acid Succinimidyl Ester/Multiwalled Carbon Nanotube and Au Nanoparticle Modified Electrode , 2008 .

[44]  Da Chen,et al.  Graphene-based materials in electrochemistry. , 2010, Chemical Society reviews.

[45]  Freddy Yin Chiang Boey,et al.  Direct Electrochemical Reduction of Single-Layer Graphene Oxide and Subsequent Functionalization with Glucose Oxidase , 2009 .

[46]  Chunhai Fan,et al.  Graphene oxide-facilitated electron transfer of metalloproteins at electrode surfaces. , 2010, Langmuir : the ACS journal of surfaces and colloids.